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Biomedical subjects

H W Liu

Publications and source records attributed to H W Liu.

At least 55 records · Page 3Linked to original sources

Intrastriatal transplantation of Sertoli cells may improve amphetamine-induced rotation and tyrosine hydroxylase immunoreactivity of the striatum in hemiparkinsonian rats.

This study investigated survival and neurotrophic effects of Sertoli cells transplanted in the striatum of 6-hydroxydopamine (6-OHDA)-induced hemiparkinsonian rats. Primary cultures of Sertoli cells were established from 3-week old rats and characterized by associated marker, placental cadherin (P-cadherin). Two months after transplantation, amphetamine-induced rotations of rats transplanted with Sertoli cells were significantly lower than those of control rats. However, restoration of tyrosine hydroxylase (TH) immunoreactivity and Sertoli cells that expressed P-cadherin were only found in the striatum of the rat that showed full recovery from amphetamine-induced rotation 3 months after transplantation without immunosuppression. These results suggest that Sertoli cells transplanted in striatum of hemiparkinsonian rats may survive for at least 3 months, and improve amphetamine-induced rotation and restore TH immunoreactivity.

Amphetamine↗

Biosynthesis of desosamine: construction of a new macrolide carrying a genetically designed sugar moiety.

[formula: see text] The appended sugars in macrolide antibiotics are indispensable to the biological activities of these important drugs. In an effort to generate a set of novel macrolide derivatives, we have created a new analogue of methymycin and neomethymycin, antibiotics produced by Streptomyces venezuelae. This analogue 15 carrying a different sugar, D-quinovose, instead of D-desosamine, was constructed by taking advantage of targeted gene deletion combined with a specific pathway-independent C-3 reduction capability of the wild type S. venezuelae.

Amino Sugars↗

Hepatitis C virus infection among short-term intravenous drug users in southern Taiwan.

The purpose of this study was to determine the correlation between the prevalence of hepatitis C virus (HCV) infection with duration of drug use and other risk factors among drug users. This survey covered 899 male drug users from Kaohsiung Narcotic Abstention Institute and Kaohsiung prison. The prevalence of positive anti-HCV was 67.2% among intravenous drug users (IVDU) and 14.7% among non-intravenous drug users (non-IVDU). Among intravenous (IV) drug users, age and duration of drug use were independently related to HCV seropositivity. Seroprevalence rate for HCV in the IVDU group increased with increasing duration of injection use within the first seven years of drug use. However, the steepest trajectory in seroprevalence of HCV infection occurred within the first four months. Due to the high rate of HCV infection among drug users, investigation of risk behaviors should be routine in such a group. A high frequency of HCV infection was also found among short-term injectors, which indicated that early risk reduction intervention was an important measure in moderating HCV infection.

Adolescent↗

Reduction of platelet transfusion- associated sepsis by short-term bacterial culture.

BACKGROUND AND OBJECTIVES: There is as yet no suitable routine laboratory test for a blood transfusion service to detect bacterial contamination in platelets. This study evaluates the effectiveness and the applicability of short-term bacterial culture for such a purpose. MATERIALS AND METHODS: Samples from 5-unit platelet pools were inoculated into an aerobic culture bottle, then monitored for 48 h at 35 degrees C in an automated monitoring and detection system. RESULTS: 26,210 whole-blood-derived platelet components were tested, of which 14 (0.053%) platelet units were found to be contaminated. In addition, nine of the associated red cell units and 4 fresh-frozen plasma units grew the same organisms on culture. CONCLUSION: Short-duration bacterial culture by an automated system is effective and suitable for routine screening in a regional transfusion center.

Colony-Forming Units Assay↗

Pulmonary immunity to Listeria is enhanced by elimination of alveolar macrophages.

To determine how resident alveolar macrophages (AM) regulate the antigen-presenting-cell (APC) activities of pulmonary dendritic cells (DC) in the response to particulate antigen, we pretreated Lewis rats intratracheally with liposomes containing clodronate (LIP-CLOD), which eliminated AM in vivo. Controls received saline encapsulated in liposomes (LIP-SAL) or saline alone intratracheally. At Day 3, rats were injected intratracheally with 1 x 10(7) heat-killed Listeria (HKL) and DC purified from lung were examined for their ability to stimulate HKL-immune T cells without added HKL. Only DC from LIP-CLOD-treated rats displayed enhanced APC activities for HKL. A second intratracheal HKL challenge at Day 14 yielded lymphocytic cuffing of the microvasculature in LIP-CLOD-treated lungs only. Intratracheal adoptive transfer of normal syngeneic AM into LIP-CLOD-treated rats suppressed APC activities of DC in vitro and the lymphocytic response in vivo. Bronchoalveolar macrophages from rats treated with LIP-CLOD and HKL showed decreased production of nitric oxide (NO), a potent suppressor of DC and T-helper 1 lymphocyte activities as compared with those of controls. We conclude that eliminating AM in vivo reduces local production of NO and promotes pulmonary cell-mediated immunity to HKL.

Animals↗

HLA-DMA and HLA-DMB genotyping in patients with rheumatic diseases.

To investigate the correlation of HLA-DMA and DMB alleles to some rheumatic diseases, HLA-DMA and DMB genes were detected in 11 patients with juvenile rheumatoid arthritis (JRA), 22 patients with psoriatic arthritis, 26 patients with Behcet's disease, 62 patients with ankylosing spondylitis (AS), and 138 unrelated healthy controls. There was no significant difference in phenotypic frequencies of HLA-DMA and DMB alleles between controls and patients with these rheumatic diseases. HLA-DMA and DMB genes are not related to the susceptibility of JRA, psoriatic arthritis, Behcet's disease, and AS.

Alleles↗

HLA-DMA and HLA-DMB genotyping in patients with systemic lupus erythematosus.

UNLABELLED: OBJECTIVE. To investigate the relation of HLA-DMA and DMB alleles to the development of systemic lupus erythematosus (SLE). METHODS: We studied HLA-DMA and DMB alleles in 108 patients with SLE and 138 healthy controls using the amplification created restriction site method. RESULTS: The phenotypic frequency of HLA-DMA*0104 was significantly higher in patients with SLE than in healthy controls. The HLA-DMA and DMB alleles were not related to clinical manifestations and various laboratory tests in patients with SLE. CONCLUSION: HLA-DMA*0104 may be associated with the susceptibility of SLE.

Adolescent↗

A gene cluster for macrolide antibiotic biosynthesis in Streptomyces venezuelae: architecture of metabolic diversity.

In a survey of microbial systems capable of generating unusual metabolite structural variability, Streptomyces venezuelae ATCC 15439 is notable in its ability to produce two distinct groups of macrolide antibiotics. Methymycin and neomethymycin are derived from the 12-membered ring macrolactone 10-deoxymethynolide, whereas narbomycin and pikromycin are derived from the 14-membered ring macrolactone, narbonolide. This report describes the cloning and characterization of the biosynthetic gene cluster for these antibiotics. Central to the cluster is a polyketide synthase locus (pikA) that encodes a six-module system comprised of four multifunctional proteins, in addition to a type II thioesterase (TEII). Immediately downstream is a set of genes for desosamine biosynthesis (des) and macrolide ring hydroxylation. The study suggests that Pik TEII plays a role in forming a metabolic branch through which polyketides of different chain length are generated, and the glycosyl transferase (encoded by desVII) has the ability to catalyze glycosylation of both the 12- and 14-membered ring macrolactones. Moreover, the pikC-encoded P450 hydroxylase provides yet another layer of structural variability by introducing regiochemical diversity into the macrolide ring systems. The data support the notion that the architecture of the pik gene cluster as well as the unusual substrate specificity of particular enzymes contributes to its ability to generate four macrolide antibiotics.

Anti-Bacterial Agents↗

Characterization of Trichomonas vaginalis virus proteins in the pathogenic protozoan T. vaginalis.

The 4.6-kb double-stranded (ds) RNA of Trichomonas vaginalis virus (TVV)-T1 has been shown to encode two overlapping genes, cap and pol. In this study, a serum for specifically detecting viral cap gene product was raised against a recombinant protein, and sera for specifically detecting pol gene product were raised against synthetic oligopeptides. A 75-kDa major protein and a 160-kDa minor protein were detected by anti-CAP serum in a TVV-T1 sample, indicating that the 75-kDa protein is the viral capsid protein. The 160-kDa protein alone was also detected by two distinct anti-POL sera, indicating that the pol gene is expressed as a CAP-POL fusion protein. These results suggest that the TVV-T1 genome is arranged into a cap-pol organization in a manner similar to that of viruses in family Totiviridae.

Animals↗

HLA-DP epitope typing using monoclonal antibodies.

We have made a panel of murine anti-DP monoclonal antibodies for serological typing of HLA-DP polymorphisms; they can be used in microcytotoxicity (for 7 epitopes) and binding assays (for 8 epitopes). The antibodies detect polymorphic differences in both alpha and beta chains. As immunogens we sometimes used B-lymphoblastoid lines or purified DP molecules but mostly used mouse fibroblast transfectants expressing DP molecules. The DP beta genes were made from a cloned DPB1*0201 gene by replacing its major area of polymorphism with matching stretches of DNA amplified from other alleles; cloned DPA1*01 and DPA1*02 genes were used for transfection along with the beta chain genes. The monoclonal antibodies showed reaction patterns that correlated with the presence of particular amino-acid sequence motifs; thus none of the antibodies is allele-specific. They bind instead to epitopes which are found on a number of different HLA-DP types. We have constructed frequency tables so that the epitope (motif) data can be interpreted as the most likely genotype in each case. The basic assumption to justify this work is that HLA-DP matching or mismatching will likely influence transplant outcome, particularly in bone marrow transplantation. The present challenge is to define permissive and nonpermissive combinations of HLA-DP; it may be that matching for epitopes, rather than for full alleles, will help to resolve this issue.

Animals↗

T cell receptor gene V alpha and V beta usage in patients with rheumatoid arthritis in Taiwan.

To investigate the correlation of T cell receptor (TCR) genes to rheumatoid arthritis (RA) in Taiwan, synovial fluid and peripheral blood were examined simultaneously in 14 patients with RA, and peripheral blood only was examined in 15 healthy controls. RNA was extracted from T cell in synovial fluid and peripheral blood, and cDNA was synthesized using the reverse transcription method. Then, TCR-V alpha and V beta gene families were determined using the polymerase chain reaction and slot blot methods. The numbers of TCR-V alpha and V beta families, especially V beta, were decreased in rheumatoid synovial fluid compared with numbers found in the peripheral blood of the same RA patients. TCR-V beta 7 and V beta 8 could be found in the synovial fluid of all patients with positive HLA-DR4. However, there was no significant difference in the numbers of TCR-V alpha and V beta families in peripheral blood of RA patients and the peripheral blood of healthy controls. In conclusion, restricted heterogeneity of TCR-V gene, especially V beta, can be found in synovial fluid of patients with RA. TCR-V beta 7 and V beta 8 may be related to the pathogenesis of HLA-DR4 positive RA patients in Taiwan.

Arthritis, Rheumatoid↗

Genetic analysis of interleukin-10 promoter region in patients with systemic lupus erythematosus in Taiwan.

Overproduction of interleukin-10 (IL-10) may play an important role in the development of systemic lupus erythematosus (SLE) or lupus nephritis. There is also a polymorphic dinucleotide repeat in the human IL-10 promoter region (IL-10PR). Our aim was to study whether or not the IL-10PR alleles contributed to the susceptibility to SLE or lupus nephritis. One hundred SLE patients and 103 healthy controls were studied for IL-10PR by PCR and electrophoretic analysis. The distribution of IL-10PR alleles, genotypes and the sum of both alleles (SBA) from different groups or subgroups were analyzed. SLE patients showed no difference in the distribution of IL-10PR alleles, genotypes and SBA, as compared to healthy controls. Lupus nephritis patients (N = 49) also showed no difference in IL-10PR alleles, genotypes and SBA, as compared to SLE patients without nephritis (N = 51). Of 49 lupus nephritis patients, ten developed end-stage renal disease (ESRD) and four of them were found to suffer from rapid progressive renal failure (RPRF). Patients with RPRF presented much smaller SBA than other ESRD patients (p = 0.005). Lupus nephritis patients carrying small SBA (< 18) suffered from a higher prevalence of RPRF than lupus nephritis patients without small SBA (50% V.S. 0%, p < 0.001, relative risk 82). Our data provide the first evidence of a strong association between IL-10PR and severe progression of lupus nephritis in human patients. In the future, a prospective genetic analysis of IL-10PR for patients with lupus nephritis is recommended. It might be helpful for physicians to identify the lupus nephritis subgroup with a high risk of developing RPRF early, because this might lead to a better therapy and prognosis for these patients.

Adult↗

Seroepidemiology of hepatitis C virus infection among drug abusers in southern Taiwan.

The purpose of this study was to determine the risk factors for hepatitis C virus (HCV) infection among drug abusers in southern Taiwan. This survey included 935 drug abusers from Kaohsiung Narcotic Abstention Institute and Kaohsiung prison. The prevalence of anti-HCV antibody was 29.1% among male drug abusers and 19.4% among female drug abusers. The seroprevalence of anti-HCV antibody was 66.4% among intravenous drug abusers and 14.4% among nonintravenous drug abusers. Intravenous drug use, a history of hepatitis, having tattoos, and age were independently related to HCV seropositivity among drug abusers. The prevalence of anti-HCV antibody concentrations significantly increased (10.8-fold) with intravenous drug abuse and with having tattoos (1.7-fold). These findings suggest that hepatitis C virus is mainly transmitted by the parenteral route among drug abusers in southern Taiwan. Due to the high rate of HCV infection among drug abusers, investigation of high-risk behavior should be routine in this group. To prevent HCV infection, emphasis on the use of sterile needles and aseptic procedures in tattooing is important in Taiwan.

Adolescent↗

Characterization of Rab3A, Rab3B and Rab3C: different biochemical properties and intracellular localization in bovine chromaffin cells.

In this study we examined the biochemical properties and subcellular localization of Rab3A, Rab3B and Rab3C in bovine adrenal chromaffin cells. The Kd for guanosine 5'-[gamma-thio]triphosphate (GTP[S]) of the three Rab3 proteins was 15, 2700 and 204 nM for Rab3A, Rab3B and Rab3C respectively. The intrinsic GTPase activity of the three Rab3 proteins seemed similar and was increased approx. 3-fold by bovine chromaffin cell lysate. Truncation of the C-terminal 31 amino acid residues decreased the binding affinity for GTP[S] of the three Rab3 proteins. When the C-terminus of Rab3C was replaced with that of Rab3A, the binding affinity of Rab3C for GTP[S] was decreased, but the replacement did not affect the affinity of Rab3B for GTP[S]. Immunostaining experiments showed that Rab3A, Rab3B and Rab3C are localized separately within chromaffin cells. Anti-Rab3A and anti-Rab3C antibodies stained vesicle-like structures, whereas anti-Rab3B antibody distinctly stained the plasma membrane. In summary, bovine chromaffin cells express the three Rab3 proteins but the subcellular localization and biochemical properties of the three Rab3 proteins are distinct.

Adrenal Medulla↗

The TNF2 allele does not contribute towards susceptibility to systemic lupus erythematosus.

The uncommon allele (TNF2) of a polymorphism in the promoter region of the tumor necrosis factor-alpha (TNF-alpha) gene has been reported to be increased in Caucasian systemic lupus erythematosus (SLE) patients (associated with HLA-DR3). To investigate whether TNF2 contributes towards susceptibility to Chinese SLE patients (not associated with HLA-DR3), 100 patients with SLE and 107 controls were studied. The frequency of TNF2 allele in controls was 0.140. There was a strong association between TNF2 allele and HLA-DR3 (P < 10(-8)) in controls. The frequency of TNF2 allele in SLE patients was 0.15. There was no difference in frequencies of TNF2 allele between patients and controls. This finding strongly suggests that TNF2 does not play a direct role in the susceptibility of SLE.

Alleles↗

Redesigning the active-site of an acyl-CoA dehydrogenase: new evidence supporting a one-base mechanism.

The acyl-CoA dehydrogenases are a family of related enzymes that share high structural homology and a common catalytic mechanism which involves abstraction of an alpha-proton from the substrate by an active site glutamate residue. Several lines of investigation have shown that the position of the catalytic glutamate is conserved in most of these dehydrogenases (the E2 site), but is in a different location in two other family members (the E1 site). Using site specific in vitro mutagenesis, a double mutant rat short chain acyl-CoA dehydrogenase (rSCAD) has been constructed in which the catalytic glutamate is moved from the E2 to the E1 site (Glu368Gly/Gly247Glu). This mutant enzyme is catalytically active, but utilizes substrate less efficiently than the native enzyme (K(m) = 0.6 and 2.0 microM, and Vmax = 2.8 and 0.3 s-1 for native and mutant enzyme respectively). In this study we show that both the wild-type and mutant rSCADs display identical stereochemical preference for catalysis--abstraction of the alpha-HR from the substrate followed by transfer of the beta-HR to the FAD coenzyme. These results, in conjunction with molecular modeling of the native and double mutant SCAD indicate that the catalytic base in the E1 and E2 sites are topologically similar and catalytically competent. However, analysis of the 1H NMR spectra of the incubation products of these two enzymes revealed that, in contrast to the wild-type rSCAD, the Gly368Glu/Gly247Glu rSCAD could not perform gamma-proton exchange of the product with the solvent, a property inherent to most acyl-CoA dehydrogenases. It is evident that the base in the mutant enzyme has access to the alpha-HR but is far removed from the gamma-Hs. These findings provide further support for a one base mechanism of alpha- and gamma-reprotonation/deprotonation catalysis by acyl-CoA dehydrogenases.

Acyl-CoA Dehydrogenases↗

Elevated serum liver enzymes in coke oven and by-product workers.

Coke oven and by-product workers are potentially exposed to coke oven emissions (COE), which contain hundreds of chemicals and are primarily composed of polycyclic aromatic hydrocarbons (PAH) and volatile organic compounds. Some of these compounds are hepatotoxins. The objective of this study was to examine the relationship between work in coke oven and by-product plants and serum activities of aspartate aminotransferase (AST) and alanine aminotransferase (ALT), the most commonly performed liver-function tests. The exposed group was composed of current workers who had been employed at least 3 months in the two coke-operation work areas, including one coke oven plant and one by-product plant (Area I: n = 117; Area II: n = 96) of a large steel company in Taiwan. Control subjects (Area III: n = 131), not visiting either coke-operation area in the last 3 months, were collected from the administrative and nonproduction areas in the same company. PAH exposure, as a surrogate of COE, was measured monthly by PM-10 size-selective high-volume-area air samplers in or around these three areas between June and December 1990, as well as between November 1992 and June 1993. The mean total respiratory particulate PAH exposure levels (< 10 microns) between November 1992 and June 1993 in Area I, II, and III were 6.8 x 10(3), 2.1 x 10(3), and 6.5 x 10(1) ng/m3, respectively. AST, ALT, and hepatitis B surface antigen tests were performed in 1994. Workers who showed either AST or ALT levels greater than reference levels (abnormal > 25 IU/L) were regarded as showing "elevated liver enzyme levels." Workers in Area I had AST levels that were 17% higher (95% confidence interval [CI], 3% to 32%]) and ALT levels that were 35% higher (95% CI, 10% to 65%)] than those in Area III after controlling for appropriate confounders. The adjusted odds ratio (Area I vs Area III) for elevated liver enzymes was 4.4 (95% CI, 1.5 to 13.4). In addition, coke oven (n = 91) and by-product workers (n = 26) from Area I had ALT levels 37% and 45% higher, respectively, compared with control subjects from Area III, after adjusting for appropriate confounders. Similar effects are also seen for AST. Workers in Area II had slightly, but not significantly, elevated AST and ALT levels. These results indicate that workers most heavily exposed to COE exhibit elevated aminotransferase levels.

Adult↗